Basic Clinical Microbiology & Pathology
Laboratory (35.213)
Day
Lab #
JULY 7
1
JULY 8
2
JULY 9
3
JULY 13
4
JULY 14
JULY 15
JULY 16
JULY 20
JULY 21
JULY 22
JULY 23
JULY 27
JULY 28
JULY 29
JULY 30
AUG 3
AUG 4
AUG 5
AUG 6
AUG 10
AUG 11
AUG 12
LABORATORY EXERCISE
Lab Introduction, Syllabus Review, Lab Guidelines, Lab Safety
The Use and Care of the Microscope
Preparation and Examination of Smears: Gram Stain
Examination of Prepared Smears
Preparation and Examination of Smears: Acid Fast Stain
Examination of Prepared Smears
SETUP: Aseptic Techniques and Isolation of Bacteria
RESULTS: Aseptic Techniques and Isolation of Bacteria
5
6
7
8
9
10
11
12
PARASITOLOGY: Protozoa and Worms
PRACTICAL LAB TEST #1: Labs 1 – 5 (6 pm)
SET UP: Integumentary System
RESULTS: Integumentary System
SET UP: Respiratory System
RESULTS: Respiratory System
SET UP: Urogenital System
RESULTS: Urogenital System
PARASITOLOGY: Tick Identification
PRACTICAL LAB TEST #2: Labs 6 – 9 (6 pm)
SET UP: Gastrointestinal System
RESULTS: Gastrointestinal System
SET UP: Susceptibility Testing – KB and MIC
RESULTS: Susceptibility Testing – KB and MIC
FUNGI: Yeast and Moulds
FUNGI: View prepared smears and Review
PRACTICAL LAB TEST #3: Labs 10 – 12 (6pm)
EMAIL: sarah_zimmerman@uml.edu
sarah.k.zimmerman@lahey.org
OFFICE HOURS: None. See me before or after
class.
LAB MANUAL: Custom UML LAB manual
available in UML bookstore only
Evaluation: 3 practical exams
MY EXPECTATIONS
• COME TO EVERY LAB
PREPARED BY
REVIEWING THE LAB
EXPERIMENT IN YOUR
MANUAL/PowerPoints
• THERE IS NO MAKE-UP!
MICROORGANISMS USED IN ALL
EXPERIMENTS
Many of the microorganisms we will use this
semester will be Biosafety Level 1 (not shown to
cause disease in humans) but several will be
Biosafety Level 2 (can cause disease in humans).
Because of this potential risk we ask that you
treat ALL bacterial cultures as if they cause
infection!
GENERAL LAB RULES
1. BE ON TIME
2. ATTEND EVERY LAB
3. NO: CELL PHONES, TABLETS, LAPTOPS
4. NO: FOOD OR DRINK
Lab Safety Guidelines
1. Absolutely NO food or drink.
2. Wear appropriate clothing (long pants) & shoes that
cover your entire foot (no Flip-flops/sandals/open-toe
shoes).
3. Wear a lab coat at all times.
4. Wash hands & clean off bench before and after lab.
5. Wear gloves & face shields when necessary.
6. Dispose of hazardous & nonhazardous materials
appropriately.
7. Report any spills or accidents immediately.
8. Do not touch your eyes, nose, or mouth while in lab.
LAB SAFETY NOTES
What you should know:
•
•
•
What to do in case of a fire or biological hazard.
Where safety equipment is located (fire alarm,
extinguishers, eyewash stations, first aid kits, and exits).
Where to properly dispose of waste (glass (SHARPS),
biological, or chemical).
PROPER TRASH DISPOSAL?
FORMS & SIGNATURES
USE AND CARE OF THE
MICROSCOPE
THE MICROSCOPE
• A device used to magnify objects that are too
small to be seen with the naked eye
Simple Microscope – one lens (like a magnifying glass)
Compound Microscope – two or more sets of lenses
Simple
Compound
MICROSCOPE TERMS
Specimen: the object that is being looked at on a slide
Ocular lens: EYEPIECE
Objective lens: lens closest to the specimen
Condenser: lens that concentrates the light for better resolution of the
specimen
Iris diaphragm: controls the amount of light
Rheostat: controls intensity of light
Course adjustment: used for focusing with low power objective lens
Fine adjustment: used for focusing with high power and oil immersion
lens
Parfocal – specimen will remain in focus as you change the objective
lenses
Resolution – the ability of the lenses to reveal fine detail
How does a microscope work?
PARTS OF A COMPOUND MICROSCOPE
MAGNIFICATION
• Compound microscopes have 3-4 objective
lenses:
– Scanning (4x), low power (10x), high power
dry)(high-dry) (40x), and oil immersion (100x)
• The ocular lens (eyepiece) has a magnification of
10x.
TOTAL MAGNIFICATION
DETERMINATION
COMPOUND MICROSCOPE
MAGNIFICATION – OIL (100X)
• OIL IMMERSION: PLACE A DROP OF IMMERSION OIL ON
THE SLIDE BEFORE THE 100X OBJECTIVE LENS IS USED.
THIS DECREASES THE BENDING OF THE LIGHT RAYS
(REFRACTION)
• NOTE: OIL MUST BE WIPED OFF THIS OBJECTIVE AND
THE SLIDE IN ORDER TO USE ANOTHER OBJECTIVE
TIPS FOR SUCCESS
1. HANDLE THE MICROSCOPE WITH CARE (BOTH HANDS).
2. CLEAN IT THOROUGHLY WITH ONLY LENS PAPER AT
THE END OF EACH CLASS.
3. DO NOT FORCE ANY OF THE MICROSCOPE’S
ADJUSTMENTS.
4. DO NOT DIS-ASSEMBLE ANYTHING – SEE ME IF YOU
HAVE ANY PROBLEMS.
5. MICROSCOPE SLIDES SHOULD BE PLACED WITH THE
COVERSLIP UP…CLOSEST TO THE OBJECTIVE.
TIPS FOR SUCCESS
6. ALWAYS BEGIN WITH THE 4X OBJECTIVE AND WORK
UP TO HIGHER MAGNIFICATION.
4X  10X 40X ADD OIL100X
7. ALWAYS PLACE YOUR SPECIMEN IN THE CENTER OF
YOUR FIELD OF VIEW BEFORE SWITCHING TO THE
NEXT HIGHER OBJECTIVE.
8. FOCUSING PROBLEM – MOVE THE SLIDE TO THE EDGE
OF THE COVERSLIP AND FOCUS ON THAT FIRST.
9. OBJECTS VIEW WILL GET BIGGER AS THE
MAGNIFICATION INCREASES.
BASIC BACTERIAL AND YEAST
MORPHOLOGIES
WHITE AND RED BLOOD CELLS